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17-658 ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - ChIP Validated Antibody and Primer Set

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17-658
25 assays  25 assays per kit, ~5μg per chromatin immunoprecipitation
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, M, MaWB, ICC, ChIP
      Description
      Catalogue Number17-658
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Acetyl-Histone H3 (Lys9) Purified - ChIP Validated Antibody and Primer Set
      OverviewAll ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
      The ChIPAb+ Acetyl-Histone H3 (Lys9) set includes the anti-acetyl-histone H3 (Lys9) antibody, a negative control antibody (purified Rabbit IgG), and qPCR primers flanking an Sp1 binding site in the human p21 (WAF1/CIP1/CDKN1A) promoter, amplifying a 105 base pair PCR product. The acetyl-histone H3 (Lys9) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of acetyl-histone H3 (Lys9) associated chromatin.
      Alternate Names
      • H3K9Ac
      • Histone H3 (acetyl K9)
      References
      Product Information
      FormatProtein A Purified
      Control
      • Included negative control antibody purified rabbit IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.
      PresentationAnti-Acetyl-Histone H3 (Lys9) (rabbit polyclonal IgG). One vial containing 125 μg of protein A purified antibody in 125 μL storage buffer containing 0.05% sodium azide and 30% glycerol.

      Normal Rabbit IgG. One vial containing 125 ug purified Rabbit IgG in 125 uL storage buffer containing 0.05% sodium azide.

      ChIP Primers p21. One vial containing 75 μL of 5 μM of each primer specific for a region of the human p21 (WAF1/CIP1/CDKN1A) promoter.
      FOR: GTG GCT CTG ATT GGC TTT CTG
      REV: CTG AAA ACA GGC AGC CCA AG
      Quality LevelMQ100
      Applications
      ApplicationThis ChIPAb+ Acetyl-Histone H3 (Lys9) Purified -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Immunocytochemistry
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from untreated or UV treated (50 J/m2, 6 hrs.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 5 μg of either a normal rabbit IgG or Anti-Acetyl-Histone H3 (Lys9) antibody and the Magna ChIP A (Cat. #17-610) Kit. Successful immunoprecipitation of acetyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers p21 flanking the human p21(WAF1/CIP1/CDKN1A) promoter (Please see figures). Data is presented as fold enrichment of normalized percent input of each IP sample relative to input treated or untreated chromatin.
      Please refer to the EZ-Magna A ChIP™ (Cat. #17-408) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.

      Western Blot Analysis:
      Acid-extracted proteins from normal HeLa cells (Lane 1) and HeLa cells treated with 5 mM sodium butyrate for 24 hours (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-Acetyl Histone H3 (Lys9) (1 μg/mL). Proteins were visualized using a goat-anti rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
      Biological Information
      ImmunogenThe acetyl-histone H3 (Lys9) purified antibody is made against a peptide (acetylated at Lys9) corresponding to amino acids 1-12 of histone H3.
      HostRabbit
      SpecificityAcetyl-Histone H3 (Lys9)
      Species Reactivity
      • Human
      • Mouse
      • Mammals
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • H3F3A
      • H3F3B
      • H3F3
      UniProt Number
      UniProt SummaryFUNCTION:Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Ref.14 Ref.18 Ref.22

      SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Interacts with HIRA, a chaperone required for its incorporation into nucleosomes. Ref.14

      SUBCELLULAR LOCATION: Nucleus.

      Developmental stage Expressed throughout the cell cycle independently of DNA synthesis.

      PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18.

      Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription.

      Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression.

      Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5, Lys-37 and Lys-80. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28, which are linked to gene repression, are underrepresented. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin.

      Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase, probably DAPK3. Phosphorylation at 'Ser-11' by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation on Ser-32 is specific to regions bordering centromeres in metaphase chromosomes. Ref.9 Ref.10 Ref.12 Ref.13 Ref.19 Ref.20 Ref.21 Ref.29

      Ubiquitinated By similarity.

      SIMILARITY: Belongs to the histone H3 family.

      SEQUENCE CAUTION: The sequence CAH73371.1 differs from that shown. Reason: Erroneous gene model prediction.

      Molecular Weight17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from UV treated (50 J/m2, 6 hrs.) U2OS cells (3 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 5 μg of either a normal rabbit IgG or Anti-Acetyl-Histone H3 (Lys9) antibody and the Magna ChIP A (Cat. #17- 610) Kit. Successful immunoprecipitation of acetyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers p21 flanking the human p21(WAF1/CIP1/CDKN1A) promoter (Please see figures).
      Please refer to the EZ-Magna A ChIP™ (Cat. #17-408) or EZ-ChIP™ (Cat. #17- 371) protocol for experimental details.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at -20°C from date of receipt
      Packaging Information
      Material Size25 assays
      Material Package25 assays per kit, ~5μg per chromatin immunoprecipitation
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ Acetyl-Histone H3 (Lys9) -2814954 2814954
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 1967265 1967265
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 1994402 1994402
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 2041596 2041596
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 2142720 2142720
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 2328394 2328394
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 3006577 3006577
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 3185808 3185808
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 3326881 3326881
      ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - 3452522 3452522

      References

      Reference overviewApplicationPub Med ID
      ZEB1-associated drug resistance in cancer cells is reversed by the class I HDAC inhibitor mocetinostat.
      Meidhof, S; Brabletz, S; Lehmann, W; Preca, BT; Mock, K; Ruh, M; Schüler, J; Berthold, M; Weber, A; Burk, U; Lübbert, M; Puhr, M; Culig, Z; Wellner, U; Keck, T; Bronsert, P; Küsters, S; Hopt, UT; Stemmler, MP; Brabletz, T
      EMBO molecular medicine  7  831-47  2015

      Show Abstract
      25872941 25872941
      The histone acetyltransferase MOF activates hypothalamic polysialylation to prevent diet-induced obesity in mice.
      Brenachot, X; Rigault, C; Nédélec, E; Laderrière, A; Khanam, T; Gouazé, A; Chaudy, S; Lemoine, A; Datiche, F; Gascuel, J; Pénicaud, L; Benani, A
      Molecular metabolism  3  619-29  2014

      Show Abstract
      25161885 25161885
      Transgenerational phenotypic and epigenetic changes in response to heat stress in Arabidopsis thaliana.
      Migicovsky, Z; Yao, Y; Kovalchuk, I
      Plant signaling & behavior  9  e27971  2014

      Show Abstract
      24513700 24513700
      Deletion of a conserved cis-element in the Ifng locus highlights the role of acute histone acetylation in modulating inducible gene transcription.
      Balasubramani, A; Winstead, CJ; Turner, H; Janowski, KM; Harbour, SN; Shibata, Y; Crawford, GE; Hatton, RD; Weaver, CT
      PLoS genetics  10  e1003969  2014

      Show Abstract
      24415943 24415943
      Depletion of sirtuin 1 (SIRT1) leads to epigenetic modifications of telomerase (TERT) gene in hepatocellular carcinoma cells.
      Zhang, B; Chen, J; Cheng, AS; Ko, BC
      PloS one  9  e84931  2014

      Show Abstract
      24416313 24416313
      Epigenetic regulation of MDR1 gene through post-translational histone modifications in prostate cancer.
      Henrique, R; Oliveira, AI; Costa, VL; Baptista, T; Martins, AT; Morais, A; Oliveira, J; Jerónimo, C
      BMC Genomics  14  898  2013

      Show Abstract
      24344919 24344919
      Rev-erb-α modulates skeletal muscle oxidative capacity by regulating mitochondrial biogenesis and autophagy.
      Woldt, E; Sebti, Y; Solt, LA; Duhem, C; Lancel, S; Eeckhoute, J; Hesselink, MK; Paquet, C; Delhaye, S; Shin, Y; Kamenecka, TM; Schaart, G; Lefebvre, P; Nevière, R; Burris, TP; Schrauwen, P; Staels, B; Duez, H
      Nature medicine  19  1039-46  2013

      Show Abstract
      Western Blotting23852339 23852339
      CREB-binding protein (CBP) regulates β-adrenoceptor (β-AR)-mediated apoptosis.
      Lee, YY; Moujalled, D; Doerflinger, M; Gangoda, L; Weston, R; Rahimi, A; de Alboran, I; Herold, M; Bouillet, P; Xu, Q; Gao, X; Du, XJ; Puthalakath, H
      Cell death and differentiation  20  941-52  2013

      Show Abstract
      Western Blotting23579242 23579242
      Valproic acid inhibits the proliferation of cancer cells by re-expressing cyclin D2.
      Witt, D; Burfeind, P; von Hardenberg, S; Opitz, L; Salinas-Riester, G; Bremmer, F; Schweyer, S; Thelen, P; Neesen, J; Kaulfuss, S
      Carcinogenesis  34  1115-24  2013

      Show Abstract
      23349020 23349020
      Histone modifications are responsible for decreased Fas expression and apoptosis resistance in fibrotic lung fibroblasts.
      Huang, SK; Scruggs, AM; Donaghy, J; Horowitz, JC; Zaslona, Z; Przybranowski, S; White, ES; Peters-Golden, M
      Cell death & disease  4  e621  2013

      Show Abstract
      Western Blotting23640463 23640463

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      Title
      Advance your Epigenetics Research (MilliporeSigma)

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      Product Families

      Categories

      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies
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      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies